Source: AUBURN UNIVERSITY submitted to NRP
ROLE OF MUSCLE STEM CELLS IN THE WOODEN BREAST CHICKEN MEAT QUALITY DEFECT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1014981
Grant No.
2018-67017-27556
Cumulative Award Amt.
$447,675.00
Proposal No.
2017-07617
Multistate No.
(N/A)
Project Start Date
Mar 1, 2018
Project End Date
Feb 28, 2022
Grant Year
2018
Program Code
[A1361]- Improving Food Quality
Recipient Organization
AUBURN UNIVERSITY
108 M. WHITE SMITH HALL
AUBURN,AL 36849
Performing Department
Poultry Science
Non Technical Summary
We aim to understand the underlying cellular and molecular mechanisms which lead to the very costly wooden breast (WB) myopathy currently plaguing the global commercial poultry industry. This severe meat quality defect has been characterized as low quality chicken breast meat that is hard to the touch, has both poor product functionality and reduced consumer acceptability which significantly reduces its value. Our preliminary data show that the muscle stem cell populations, which are essential to muscle growth, are altered in WB-affected muscles. We propose to use both in vivo and in vitro approaches to investigate whether muscle stem cell malfunction leads to the development of this severe chicken meat quality defect. The fact that the cause of this costly meat quality defect is currently unknown is the rationale for this work.
Animal Health Component
25%
Research Effort Categories
Basic
75%
Applied
25%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3023299102025%
3053299102025%
3023299103025%
3053299103025%
Knowledge Area
302 - Nutrient Utilization in Animals; 305 - Animal Physiological Processes;

Subject Of Investigation
3299 - Poultry, general/other;

Field Of Science
1030 - Cellular biology; 1020 - Physiology;
Goals / Objectives
The overall goal of this project is to understand the underlying cellular and molecular mechanisms which lead to the very costly wooden breast (WB) myopathy currently plaguing the global commercial poultry industry. Our specific project objectives are to: 1) Determine how the size, heterogeneity, and mitotic activity of the muscle stem cell populations are impacted over time by varying WB severity, 2) Determine whether muscle stem cell function is impaired in WB-affected broilers, and 3) Characterize the changes in major muscle stem cell signaling protein pathways in muscle tissue and muscle stem cell cultures from broilers affected by varying degrees of WB over time.
Project Methods
We propose to use both in vivo and in vitro approaches to investigate whether muscle stem cell malfunction leads to the development of this severe chicken meat quality defect.

Progress 03/01/18 to 02/28/22

Outputs
Target Audience:Work this period benefited commercial poultry meat producers in developing approaches to maximize the efficiency of lean meat production as well as aid them in developing selection tools and management strategies to eliminate the Wooden Breast quality defect from the chicken meat supply. This work also benefited other researchers by increasing our knowledge of the cellular and molecular mechanisms regulating skeletal muscle growth. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Twelve graduate research assistants, 1 technician, 6 undergraduate research assistants, and 10 visiting research scholars all received training in the various animal husbandry, sample collection, lab techniques, data analysis, and oral/poster presentation of data, both in-person and virtually. Students at all levels presented at local and international scientific conferences and symposia and interacted with other scientists and industry stakeholders. How have the results been disseminated to communities of interest?The results from this project reporting period have been presented at state, national, and international scientific meetings where industry stakeholders and other research scientists were in attendance. Results were published in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? To meet the huge global demand for chicken meat, the commercial broiler industry has placed tremendous genetic selection pressure on breast meat yield, growth rate, and feed efficiency traits and has made remarkable improvements over the last 30 years. Unfortunately, along with those tremendous improvements has come a severe meat quality defect whose cause has yet to be elucidated. The Wooden (or Woody) Breast (WB) meat quality defect is characterized by visible bulging of the breast fillet with extreme hardness to the touch. The WB phenotype has been characterized by histopathologists as a degenerative myopathy in which excessive fibrotic tissue infiltration occurs. Skeletal muscle stem cells also called satellite cells (MSC) play a critical role in post-hatch broiler skeletal muscle growth, repair, and maintenance. Yet, the relationship between MSC function in high-yielding broilers and the development of the WB myopathy is still not well understood. Objective 1. Our work using in vivo cell labeling, cryohistology, and immunofluorescence techniques demonstrates that the relative size and mitotic activity of the various MSC and macrophage populations and collagen deposition are altered in WB-affected muscle compared with normal. WB-affected chickens have more proliferating MSC, greater macrophage density, and increased collagen deposition compared with un-affected birds. Our work in this area has increased scientific knowledge of the cellular and molecular characteristics of WB-affected broiler muscle and provides a starting point for further investigation. Objective 2. We have successfully isolated and cultured MSC from WB-affected and unaffected broilers and have characterized their regulatory factor expression heterogeneity over time in culture. In addition, the culture conditions for MSC from today's high-yielding broilers, including temperature, plating strata, oxygen and carbon dioxide concentrations as well as media composition have all been assessed. We found that MSC from modern, high-yielding broilers proliferate and differentiate effectively when cultured in warmer temperatures closer to normal bird body temperature and with lower oxygen concentrations. The MSC can be grown and differentiate into myotubes on gelatin-coated culture dishes. We have also established a broiler rearing model using starter diet nutrient restriction with slows growth rate and with which we can reliably produce both normal and WB-affected broilers to further study the etiology of this costly meat quality defect. Objective 3. Alterations in myogenic regulatory factor, collagen, and pro- and anti-inflammatory cytokine protein expression in WB-affected muscles have also been observed using quantitative fluorescent Western blotting. These data suggest the involvement of aberrant MSC function in the development of WB myopathy. The accomplishments from this project have increased knowledge of scientists, trainees, poultry producers, and consumers regarding the role of MSC in the WB chicken meat quality defect. The results from this work provide a good starting point for further exploration at the cellular and molecular level, including protein sequencing. Further exploration will be required to understand how the local cell signaling mechanisms and cell population kinetics are related to the severity and timing of the development of the WB myopathy in today's fast-growing, high-yielding broilers. The results of this work are useful for industry stakeholders and research scientists in determining potential interventions to reduce the incidence of or eliminate this costly meat quality defect from the chicken meat industry.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Wall, B. L., J. J. Flees, C. R. Gregg, and J. D. Starkey. 2021. Effect of cell culture plate gelatin coating method on density of primary broiler chicken skeletal muscle satellite cells. Poult. Sci. 100 (E-suppl.)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Flees, J. J., B. L. Wall, C. R. Gregg, and J. D. Starkey. 2021. Effect of basal plating culture media on primary broiler chicken muscle satellite cell myogenic regulatory factor expression heterogeneity. Poult. Sci. 100 (E-suppl.)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Gregg, C. R., J. J. Flees, B. L. Wall, and J. D. Starkey. 2021. Effect of in vitro culture temperature on heterogeneity of myogenic regulatory factor expression in primary broiler chicken satellite cells. Poult. Sci. 100 (E-suppl.)


Progress 03/01/20 to 02/28/21

Outputs
Target Audience:Work this period benefited commercial poultry meat producers in developing approaches to maximize the efficiency of lean meat production as well as aid them in developing selection tools and management strategies to eliminate the Wooden Breast quality defect from the chicken meat supply. This work also benefited other researchers by increasing our knowledge of the cellular and molecular mechanisms regulating skeletal muscle growth. Changes/Problems:The unexpected COVID pandemic and subsequent disruption in normal research operations caused significant delays in our research project progress during this reporting period, particularly for objectives 2 and 3. The objectives still remain the same and we expect to complete the project in the next reporting period. What opportunities for training and professional development has the project provided?Eight graduate research assistants, 10 undergraduate research assistants, and 7 visiting research scholars all received training in the various animal husbandry, sample collection, lab techniques, data analysis, and oral/poster presentation of data, both in-person and virtually. Students at all levels presented at local and international scientific conferences and symposia and interacted with other scientists and industry stakeholders. How have the results been disseminated to communities of interest?The results from this project reporting period have been presented at state, national, and international scientific meetings where industry stakeholders and other research scientists were in attendance. Results were published in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? To meet the huge global demand for chicken meat, the commercial broiler industry has placed tremendous genetic selection pressure on breast meat yield, growth rate, and feed efficiency traits and has made remarkable improvements over the last 30 years. Unfortunately, along with those tremendous improvements has come a severe meat quality defect whose cause has yet to be elucidated. The Wooden (or Woody) Breast (WB) meat quality defect is characterized by visible bulging of the breast fillet with extreme hardness to the touch. The WB phenotype has been characterized by histopathologists as a degenerative myopathy in which excessive fibrotic tissue infiltration occurs. Skeletal muscle stem cells also called satellite cells (MSC) play a critical role in post-hatch broiler skeletal muscle growth, repair, and maintenance. Yet, the relationship between MSC function in high-yielding broilers and the development of the WB myopathy is still not well understood. Our recent work using in vivo cell labeling, cryohistology, and immunofluorescence techniques demonstrates that the relative size and mitotic activity of the various MSC and macrophage populations and collagen deposition are altered in WB-affected muscle compared with normal. Alterations in myogenic regulatory factor, collagen, and pro- and anti-inflammatory cytokine protein expression in WB-affected muscles have also been observed using quantitative fluorescent Western blotting. These data suggest the involvement of aberrant MSC function in the development of WB myopathy. Further exploration will also be required to understand how the local cell signaling mechanisms and cell population kinetics are related to the severity and timing of the development of the WB myopathy in today's fast-growing, high-yielding broilers. The results of this work are useful for industry stakeholders and research scientists in determining potential interventions to reduce the incidence of or eliminate this costly meat quality defect from the chicken meat industry

Publications

  • Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Luis Avila. 2020. Effect of maternal and post-hatch dietary 25-hydroxycholecalciferol supplementation on broiler chicken growth characteristics and skeletal muscle satellite cell activity. Auburn University MS Thesis.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Tejeda, O. J., K. J. Meloche, J. D. Starkey. 2020. Effect of incubator tray location on broiler chicken growth performance, carcass part yields, and the meat quality defects wooden breast and white striping. Poult. Sci. 100:654-662. https://doi.org/10.1016/j. psj.2020.10.035
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Starkey, J. D. 2020. Role of skeletal muscle satellite cells in the broiler chicken Wooden Breast meat quality defect. 2020 American Society of Animal Science Virtual Meat and Muscle Biology Symposium. J. Anim. Sci. 98 (E-Suppl 1.)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Avila, L. P., S. F. Leiva, K. Sweeney, J. L. Wilson, T. Pokoo-Aikins, G. Litta, C. W. Starkey, and J. D. Starkey. 2020. Effect of maternal and post-hatch dietary 25-hydroxycholecalciferol on broiler chicken growth performance and carcass characteristics. Poult. Sci. 99 (E-Suppl 1).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Avila, L. P., S. F. Leiva, G. A. Abascal-Ponciano, J. J. Flees, K. M. Sweeney, J. L. Wilson, A. Pokoo-Aikins, S.P. Corray, G. Litta, C. W. Starkey, and J. D. Starkey. 2020. Effect of combined maternal and post-hatch dietary 25- hydroxycholecalciferol supplementation on broiler chicken Pectoralis major muscle growth characteristics and Wooden Breast. Poult. Sci. 99 (E-Suppl 1).


Progress 03/01/19 to 02/29/20

Outputs
Target Audience:This work will benefit commercial poultry meat producers in developing approaches to maximize the efficiency of lean meat production as well as aid them in developing selection tools and management strategies to eliminate the Wooden Breast quality defect from the chicken meat supply. This work will also benefit other researchers by increasing our knowledge of the cellular and molecular mechanisms regulating skeletal muscle growth. This work will ultimately benefit the consumers of poultry meat by contributing to the poultry industry's ability to provide an inexpensive and high-quality protein source. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The work conducted in this reporting period provided opportunities for undergraduate students, graduate students, and research assistants to gain experience and skills related to live bird husbandry, muscle sampling procedures, muscle stem cell culture as well as muscle cryohistology, immunofluorescence staining, and digital photomicroscopy and image analysis. How have the results been disseminated to communities of interest?The results from this project reporting period have been presented at state, national and international scientific meetings where industry stakeholders and other research scientists were in attendance. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we will complete the muscle stem cell isolations and assessment of in vitro muscle stem cell function assays which will allow us to complete the remaining project objectives.

Impacts
What was accomplished under these goals? Progress toward understanding how muscle stem cell function both in the live bird and in muscle stem cell cultures (objectives 1 and 2) was made in this reporting period. The initial experiments conducted in this reporting period by our team (8 graduate students, 12 undergraduate students, 1 research assistant, and the PD and co-PD) have resulted in information necessary to understand how stem cell function may be altered in broiler chickens with the wooden breast myopathy as well as how the stem cell populations in affected birds are impacted. This work resulted in data presented to poultry industry stakeholders and other research scientists at professional scientific meetings during the reporting period.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ferreira, T. Z., L. Kindlein, J. J. Flees, L. K. Shortnacy, S. L. Vieira, V. P. Nascimento, K. J. Meloche, and J. D. Starkey. 2020. Characterization of pectoralis major muscle satellite cell population heterogeneity, macrophage density, and collagen infiltration in broiler chickens affected by Wooden Breast. Front. Physiol. Invited. 11:529. doi: 10.33889/fphys.2020.00529. (80%)
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Tejeda, O. J., A. J. Calderon, J. A. Arana, K. J. Meloche, and J. D. Starkey. 2019. Broiler chicken myofiber morphometrics and myogenic stem cell population heterogeneity. Poult. Sci. 98:4123-4130.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Leiva, S. F., J. L. Sandoval, A. J. Keel, O. J. Tejeda, C. W. Starkey, and J. D. Starkey. 2020. Effect of dietary protein source on broiler chicken growth performance, myogenic stem cell activity and heterogeneity, and muscle fiber morphometrics. Poult. Sci. 99 (E-Suppl 1).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Flees, J. J., C. V. McGuire Sams, and J. D. Starkey. 2019. Effect of different basal culture media and sera combinations on primary broiler chicken muscle satellite cell differentiation and heterogeneity. Poult. Sci. 98 (E-Suppl 1).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Avila, L. P., S. F. Leiva, G. A. Abascal-Ponciano, J. J. Flees, K. M. Sweeney, J. L. Wilson, A. Pokoo-Aikins, S.P. Corray, G. Litta, C. W. Starkey, and J. D. Starkey. 2020. Effect of combined maternal and post-hatch dietary 25-hydroxycholecalciferol supplementation on broiler chicken Pectoralis major muscle growth characteristics and Wooden Breast. Poult. Sci. 99 (E-Suppl 1).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Flees, J. J., A. J. Keel, C. W. Starkey, and J. D. Starkey. 2020. Effect of light intensity and reduction of dietary energy and digestible lysine density on broiler chicken growth performance, breast meat yield, and meat quality defects. Poult. Sci. 99 (E-Suppl 1).


Progress 03/01/18 to 02/28/19

Outputs
Target Audience:This work will benefit commercial poultry meat producers in developing approaches to maximize the efficiency of lean meat production as well as aid them in developing selection tools and management strategies to eliminate the Wooden Breast quality defect from the chicken meat supply. This work will also benefit other researchers by increasing our knowledge of the cellular and molecular mechanisms regulating skeletal muscle growth. This work will ultimately benefit the consumers of poultry meat by contributing to the poultry industry's ability to provide an inexpensive and high quality protein source. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The work conducted in this reporting period provided opportunities for undergraduate students, graduate students, and research assistants to gain experience and skills related to muscle stem cell culture as well as muscle cryohistology, immunofluorescence staining, and digital photomicroscopy and image analysis. How have the results been disseminated to communities of interest?The results from this project reporting period have been presented at state, national and international scientific meetings where industry stakeholders and other research scientists were in attendance. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we will work toward completing the muscle stem cell isolations and assessment of in vitro muscle stem cell function assays which will allow us to complete the majority of the project objectives.

Impacts
What was accomplished under these goals? Progress toward understanding how muscle stem cell function both in the live bird and in muscle stem cell cultures (objectives 1 and 2) was made in this reporting period. The initial experiments conducted in this reporting period by our team (2 graduate students, 6 undergraduate students, 1 research assistant and the PD) have resulted in information necessary to understand how stem cell function may be altered in broiler chickens with the wooden breast myopathy as well as how the stem cell populations in affected birds are impacted. This work resulted in data presented to poultry industry stakeholders and other research scientists at professional scientific meetings during the reporting period.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Ferreira, T. Z., L. Kindlein, K. J. Meloche, S. Vieira, V. Nascimento, and J. D. Starkey. 2018. Characterization of myogenic stem cell populations in broiler chickens affected with the Wooden Breast myopathy. Poult. Sci. Vol. 97 (E-Suppl 1).
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Meloche, K. J., W.A. Dozier, III, and J. D. Starkey. 2018. Skeletal muscle growth characteristics and myogenic stem cell activity in broiler chickens affected by wooden breast. Poult. Sci. 97:4401-4414.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Flees, J. J., C. V. McGuire Sams, and J. D. Starkey. 2019. Effect of different basal culture media and sera combinations on primary broiler chicken muscle satellite cell proliferation and heterogeneity. Poult. Sci. 98 (E-Suppl 1).